Generating Isoform-Specific Antibodies: Lessons from Nucleocytoplasmic Glycoprotein Skp1
| Autor: | West, Christopher M., Van Der Wel, Hanke, Chinoy, Zoiesha, Boons, Geert Jan, Gauthier, Ted J., Taylor, Carol M., Xu, Yuechi, Medicinal Chemistry and Chemical Biology, Sub Chemical pharmacology |
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| Přispěvatelé: | Medicinal Chemistry and Chemical Biology, Sub Chemical pharmacology |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Gene isoform
Monoclonal antibody medicine.drug_class Peptide Biochemistry 03 medical and health sciences medicine Isoform-specific antibody 030304 developmental biology Polyclonal antibody chemistry.chemical_classification Medicine(all) 0303 health sciences biology Biochemistry Genetics and Molecular Biology(all) 030302 biochemistry & molecular biology Primary and secondary antibodies Molecular biology Cytoplasmic glycosylation 3. Good health Blot Oxygen Hydroxyproline Ubiquitin ligase chemistry Polyclonal antibodies biology.protein Synthetic glycopeptides Skp1 Glycoprotein Conformational epitope Genetics and Molecular Biology(all) |
| Zdroj: | Glycoscience: Biology and Medicine ISBN: 9784431548409 Glycoscience: Biology and Medicine, 927. Springer Japan STARTPAGE=927;TITLE=Glycoscience: Biology and Medicine |
| DOI: | 10.1007/978-4-431-54841-6_170 |
| Popis: | Antibodies that discriminate protein isoforms differing by modifications at specific amino acids have revolutionized studies of their functions. Skp1 is a novel nucleocytoplasmic glycoprotein that is hydroxylated at proline-143 and then O-glycosylated by a pentasaccharide attached via a GlcNAcα1, 4(trans)- hydroxyproline linkage. Skp1 isoform-specific antibodies were successfully obtained by immunizing mice or rabbits with KLH-coupled synthetic peptides bearing either unmodified Pro, 4(trans)-hydroxyproline, or D-GlcNAcα1,4 (trans)-hydroxyproline, and screening with corresponding BSA-conjugates or by Western blotting toward a panel of Skp1 isoforms. Antibodies specific for Skp1 or HO-Skp1 were not found in exhaustive murine trials, yet monospecific polyclonal antibodies were readily achieved in rabbits without crossadsorption. In all cases, antibodies were specific at the protein but not the peptide level, which suggests that conformation comprises part of the basis for recognition and which should be considered when developing screening strategies. |
| Databáze: | OpenAIRE |
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